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Lannig, Gisela; Eilers, Silke; Pörtner, Hans-Otto; Sokolova, Inna A; Bock, Christian (2010): Seawater carbonate chemistry and biological processes of mussel Crassostrea gigas during experiments, 2011. PANGAEA, https://doi.org/10.1594/PANGAEA.761915, Supplement to: Lannig, G et al. (2010): Impact of ocean acidification on energy metabolism of oyster, Crassostrea gigas—Changes in metabolic pathways and thermal response. Marine Drugs, 8(8), 2318-2339, https://doi.org/10.3390/md8082318

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Abstract:
Climate change with increasing temperature and ocean acidification (OA) poses risks for marine ecosystems. According to Pörtner and Farrell [1], synergistic effects of elevated temperature and CO2-induced OA on energy metabolism will narrow the thermal tolerance window of marine ectothermal animals. To test this hypothesis, we investigated the effect of an acute temperature rise on energy metabolism of the oyster, Crassostrea gigas chronically exposed to elevated CO2 levels (partial pressure of CO2 in the seawater ~0.15 kPa, seawater pH ~ 7.7). Within one month of incubation at elevated PCO2 and 15 °C hemolymph pH fell (pHe = 7.1 ± 0.2 (CO2-group) vs. 7.6 ± 0.1 (control)) and PeCO2 values in hemolymph increased (0.5 ± 0.2 kPa (CO2-group) vs. 0.2 ± 0.04 kPa (control)). Slightly but significantly elevated bicarbonate concentrations in the hemolymph of CO2-incubated oysters ([HCO-3]e = 1.8 ± 0.3 mM (CO2-group) vs. 1.3 ± 0.1 mM (control)) indicate only minimal regulation of extracellular acid-base status. At the acclimation temperature of 15 °C the OA-induced decrease in pHe did not lead to metabolic depression in oysters as standard metabolism rates (SMR) of CO2-exposed oysters were similar to controls. Upon acute warming SMR rose in both groups, but displayed a stronger increase in the CO2-incubated group. Investigation in isolated gill cells revealed a similar temperature-dependence of respiration between groups. Furthermore, the fraction of cellular energy demand for ion regulation via Na+/K+-ATPase was not affected by chronic hypercapnia or temperature. Metabolic profiling using 1H-NMR spectroscopy revealed substantial changes in some tissues following OA exposure at 15 °C. In mantle tissue alanine and ATP levels decreased significantly whereas an increase in succinate levels was observed in gill tissue. These findings suggest shifts in metabolic pathways following OA-exposure. Our study confirms that OA affects energy metabolism in oysters and suggests that climate change may affect populations of sessile coastal invertebrates such as mollusks
Keyword(s):
Acid-base regulation; Animalia; Benthic animals; Benthos; Biomass/Abundance/Elemental composition; Bottles or small containers/Aquaria (<20 L); Coast and continental shelf; Crassostrea gigas; Laboratory experiment; Mollusca; North Atlantic; Respiration; Single species; Temperate; Temperature
Funding:
Seventh Framework Programme (FP7), grant/award no. 211384: European Project on Ocean Acidification
Sixth Framework Programme (FP6), grant/award no. 511106: European network of excellence for Ocean Ecosystems Analysis
Comment:
In order to allow full comparability with other ocean acidification data sets, the R package seacarb (Lavigne and Gattuso, 2011) was used to compute a complete and consistent set of carbonate system variables, as described by Nisumaa et al. (2010). In this dataset the original values were archived in addition with the recalculated parameters (see related PI).
Parameter(s):
#NameShort NameUnitPrincipal InvestigatorMethod/DeviceComment
Experimental treatmentExp treatLannig, Gisela
SalinitySalLannig, Gisela
Salinity, standard deviationSal std dev±Lannig, Gisela
ReplicatesRepl#Lannig, Gisela
Temperature, waterTemp°CLannig, Gisela
pHpHLannig, GiselaMeasuredNBS scale
pH, standard deviationpH std dev±Lannig, GiselaNBS scale
Carbon dioxide, partial pressurepCO2PaLannig, GiselaCalculated using CO2SYS
Carbon dioxide, partial pressure, standard deviationpCO2 std dev±Lannig, Gisela
10 Bicarbonate ion[HCO3]-µmol/kgLannig, GiselaCalculated using CO2SYS
11 Bicarbonate ion, standard deviation[HCO3]- std dev±Lannig, Gisela
12 Calcite saturation stateOmega CalLannig, GiselaCalculated using CO2SYS
13 Calcite saturation state, standard deviationOmega Cal std dev±Lannig, Gisela
14 Aragonite saturation stateOmega ArgLannig, GiselaCalculated using CO2SYS
15 Aragonite saturation state, standard deviationOmega Arg std dev±Lannig, Gisela
16 Crassostrea gigas, haemolymph, pHC. gigas pH (ha)Lannig, Gisela
17 Crassostrea gigas, haemolymph, partial pressure of carbon dioxideC. gigas pCO2 (ha)kPaLannig, GiselaGas chromatography
18 Crassostrea gigas, haemolymph, partial pressure of oxygenC. gigas pO2 (ha)kPaLannig, GiselaGas chromatography
19 Crassostrea gigas, haemolymph, dissolved inorganic carbonC. gigas DIC (ha)mmol/lLannig, GiselaIon chromatography
20 Crassostrea gigas, haemolymph, bicarbonate ionC. gigas [HCO3]- (ha)mmol/lLannig, GiselaIon chromatography
21 Crassostrea gigas, haemolymph, calcium ionC. gigas Ca2+ (ha)mmol/lLannig, GiselaIon chromatography
22 Crassostrea gigas, haemolymph, sodium ionC. gigas Na+ (ha)mmol/lLannig, GiselaIon chromatography
23 Crassostrea gigas, haemolymph, potassium ionC. gigas K+ (ha)mmol/lLannig, GiselaIon chromatography
24 Crassostrea gigas, alanine, wet tissue massC. gigas alanineµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Muscle
25 Crassostrea gigas, succinate, wet tissue massC. gigas succinateµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Muscle
26 Crassostrea gigas, glycogen, wet tissue massC. gigas GLYµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Muscle
27 Crassostrea gigas, ATP, wet tissue massC. gigas ATPµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Muscle
28 Crassostrea gigas, alanine, wet tissue massC. gigas alanineµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Mantle
29 Crassostrea gigas, succinate, wet tissue massC. gigas succinateµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Mantle
30 Crassostrea gigas, glycogen, wet tissue massC. gigas GLYµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Mantle
31 Crassostrea gigas, ATP, wet tissue massC. gigas ATPµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Mantle
32 Crassostrea gigas, alanine, wet tissue massC. gigas alanineµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Gills
33 Crassostrea gigas, succinate, wet tissue massC. gigas succinateµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Gills
34 Crassostrea gigas, glycogen, wet tissue massC. gigas GLYµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Gills
35 Crassostrea gigas, ATP, wet tissue massC. gigas ATPµmol/gLannig, GiselaTopSpin 2.5 (Bruker Biospin GmbH, Germany)Gills
36 Condition indexCILannig, Gisela
37 Metabolic rate of oxygen, standard, normalizedSMR O2 normµmol/h/gLannig, Gisela
38 Crassostrea gigas, respiration rate, oxygen, per cellC. gigas resp O2/cellnmol/min/106#Lannig, Gisela
39 Partial pressure of carbon dioxide (water) at sea surface temperature (wet air)pCO2water_SST_wetµatmLannig, Gisela
40 Carbonate system computation flagCSC flagNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
41 pHpHNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)Total scale
42 Carbon dioxideCO2µmol/kgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
43 Partial pressure of carbon dioxide (water) at sea surface temperature (wet air)pCO2water_SST_wetµatmNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
44 Fugacity of carbon dioxide (water) at sea surface temperature (wet air)fCO2water_SST_wetµatmNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
45 Bicarbonate ion[HCO3]-µmol/kgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
46 Carbonate ion[CO3]2-µmol/kgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
47 Carbon, inorganic, dissolvedDICµmol/kgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
48 Alkalinity, totalATµmol/kgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
49 Aragonite saturation stateOmega ArgNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
50 Calcite saturation stateOmega CalNisumaa, Anne-MarinCalculated using seacarb after Nisumaa et al. (2010)
Status:
Curation Level: Enhanced curation (CurationLevelC)
Size:
2179 data points

Data

Download dataset as tab-delimited text — use the following character encoding:


Exp treat

Sal

Sal std dev [±]

Repl [#]

Temp [°C]

pH
(NBS scale, Measured)

pH std dev [±]
(NBS scale)

pCO2 [Pa]
(Calculated using CO2SYS)

pCO2 std dev [±]
10 
[HCO3]- [µmol/kg]
(Calculated using CO2SYS)
11 
[HCO3]- std dev [±]
12 
Omega Cal
(Calculated using CO2SYS)
13 
Omega Cal std dev [±]
14 
Omega Arg
(Calculated using CO2SYS)
15 
Omega Arg std dev [±]
16 
C. gigas pH (ha)
17 
C. gigas pCO2 (ha) [kPa]
(Gas chromatography)
18 
C. gigas pO2 (ha) [kPa]
(Gas chromatography)
19 
C. gigas DIC (ha) [mmol/l]
(Ion chromatography)
20 
C. gigas [HCO3]- (ha) [mmol/l]
(Ion chromatography)
21 
C. gigas Ca2+ (ha) [mmol/l]
(Ion chromatography)
22 
C. gigas Na+ (ha) [mmol/l]
(Ion chromatography)
23 
C. gigas K+ (ha) [mmol/l]
(Ion chromatography)
24 
C. gigas alanine [µmol/g]
(Muscle, TopSpin 2.5 (Bruker B...)
25 
C. gigas succinate [µmol/g]
(Muscle, TopSpin 2.5 (Bruker B...)
26 
C. gigas GLY [µmol/g]
(Muscle, TopSpin 2.5 (Bruker B...)
27 
C. gigas ATP [µmol/g]
(Muscle, TopSpin 2.5 (Bruker B...)
28 
C. gigas alanine [µmol/g]
(Mantle, TopSpin 2.5 (Bruker B...)
29 
C. gigas succinate [µmol/g]
(Mantle, TopSpin 2.5 (Bruker B...)
30 
C. gigas GLY [µmol/g]
(Mantle, TopSpin 2.5 (Bruker B...)
31 
C. gigas ATP [µmol/g]
(Mantle, TopSpin 2.5 (Bruker B...)
32 
C. gigas alanine [µmol/g]
(Gills, TopSpin 2.5 (Bruker Bi...)
33 
C. gigas succinate [µmol/g]
(Gills, TopSpin 2.5 (Bruker Bi...)
34 
C. gigas GLY [µmol/g]
(Gills, TopSpin 2.5 (Bruker Bi...)
35 
C. gigas ATP [µmol/g]
(Gills, TopSpin 2.5 (Bruker Bi...)
36 
CI
37 
SMR O2 norm [µmol/h/g]
38 
C. gigas resp O2/cell [nmol/min/106#]
39 
pCO2water_SST_wet [µatm]
40 
CSC flag
(Calculated using seacarb afte...)
41 
pH
(Total scale, Calculated using...)
42 
CO2 [µmol/kg]
(Calculated using seacarb afte...)
43 
pCO2water_SST_wet [µatm]
(Calculated using seacarb afte...)
44 
fCO2water_SST_wet [µatm]
(Calculated using seacarb afte...)
45 
[HCO3]- [µmol/kg]
(Calculated using seacarb afte...)
46 
[CO3]2- [µmol/kg]
(Calculated using seacarb afte...)
47 
DIC [µmol/kg]
(Calculated using seacarb afte...)
48 
AT [µmol/kg]
(Calculated using seacarb afte...)
49 
Omega Arg
(Calculated using seacarb afte...)
50 
Omega Cal
(Calculated using seacarb afte...)
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.7260.128718.7711.6371.2408.11439.6311.666.28220.79400.98511.25401.66930.06138.00981.21600.20820.45530.16210.27674.5623.4370.403578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.7210.15318.5921.7181.3156.69416.0410.1510.12630.86791.93301.84361.94440.30833.75831.76270.52440.03730.36300.32803.0326.2510.515578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.4980.160914.7001.5451.1506.51436.2112.326.37590.77121.18491.25223.37240.15475.35601.37200.47690.13670.78130.33603.2223.9660.473578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6530.15086.5601.7231.3366.34435.0512.429.73650.85920.63171.53351.94020.06605.70861.02800.71710.16600.80710.34733.4116.4990.239578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.7080.13856.8731.7341.3446.99437.7412.154.09850.91270.40340.35960.85180.09232.55690.81070.04620.15830.22180.35533.2614.7840.234578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6330.165712.8741.5371.1387.10446.9411.266.42150.38061.40061.73591.97870.11837.58090.77130.21780.04530.68540.350012.5900.172578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6770.16388.8381.8251.4237.22452.6411.290.15840.48730.67580.2833578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6370.20966.7927.05455.9811.680.1613578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.5090.244611.3757.77461.0112.75578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.5790.116012.9347.69472.9512.87578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6840.084013.140578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.5300.15137.966578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.3850.189414.277578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.4320.183412.648578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6100.099815.195578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511158.070.0458.638.062089.8488.323.590.442.310.307.6460.1167578.680297.9521.91577.52575.462193.52136.172351.62522.12.113.31
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3035.859578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3031.666578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3044.089578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3035.035578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3026.083578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511208.070.0458.638.062089.8488.323.590.442.310.3032.602578.680297.9418.95577.69575.752072.02151.832242.82438.62.393.71
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3046.3890.812578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3057.4580.671578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3054.0520.405578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3041.5790.340578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3042.9890.572578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Control32.120.4511258.070.0458.638.062089.8488.323.590.442.310.3040.8110.452578.680297.9316.61578.13576.311960.19168.602145.42367.72.724.15
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.2200.313211.3602.4782.0955.98415.8314.294.71790.85330.88981.74390.41420.31673.36110.33330.23930.80500.16280.27731.9321.8030.3201494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0900.57656.5741.5501.1926.26435.0213.275.14620.91821.05571.86550.07360.10676.77261.08200.19731.24430.27770.34002.7630.8540.2741494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.1700.48397.2462.1091.7526.29425.0211.432.90780.71281.00640.39430.30730.17902.01120.57530.31131.63500.65780.28733.9525.6900.2631494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0400.64269.5842.2221.8516.13420.9113.267.26300.84971.07601.31790.18180.29904.17570.65930.14070.95230.40410.36202.7923.9060.4141494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.9900.64367.0352.3281.9446.23422.3212.396.72571.10800.66381.34710.21640.07031.88200.31130.22510.33770.18730.42132.5319.7850.5111494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.2000.45819.6942.1081.7386.16414.6013.396.59610.87341.27741.73590.29420.23904.98330.30800.30730.39900.08790.28132.4321.9650.2361494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0400.65285.2362.2691.9101.28730.25006.12370.72070.27530.5281494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.2500.406211.5310.24690.04401.51420.33000.5291494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.1700.30219.9341494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.2800.368112.4321494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.3500.370410.1571494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0500.557911.6171494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.6900.88807.7751494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.9200.721511.9821494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.2600.312410.8891494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0500.47548.2061494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.3140.33667.6351494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.8120.739110.8131494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.8030.91154.7431494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0600.555410.9911494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.3250.294212.3751494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.157.0450.63799.6491494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737157.680.07151.4626.082236.70116.041.360.240.870.156.8480.83681494.760297.5656.911493.411488.092271.8455.562384.32406.70.861.36
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1541.1231494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1542.9781494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1547.1201494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1557.6391494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1559.3871494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737207.680.07151.4626.082236.70116.041.360.240.870.1563.7821494.760297.5549.211493.431488.432146.5862.012257.82297.40.981.52
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1580.4650.4691494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1573.1670.4331494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1565.9110.2471494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1571.8800.4731494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1584.7670.4901494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.1579.7460.3971494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.150.8951494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71
Incubation31.310.3737257.680.07151.4626.082236.70116.041.360.240.870.150.6811494.760297.5343.121494.551489.852031.5668.922143.62199.61.121.71